| In vitro Chondrogenic Hypertrophy Induction of Mesenchymal Stem Cells
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Author:
Date:
2016-12-05
[Abstract] To investigate underlying mechanism of chondrogenic hypertrophy, we need proper in vitro hypertrophic model of mesenchymal stem cells (MSCs). This protocol describes our defined method for induction of in vitro chondrogenic hypertrophy of human umbilical cord blood-derived MSCs (hUCB-MSCs). By adding thyroid hormone (T3; triiodothyronine) and minimum osteogenic-inducing factors to culture medium, we could induce hypertrophy of hUCB-MSCs in vitro. Hypertrophic induction was validated using immunohistochemical analysis, Western blotting and reverse transcriptase polymerase chain reaction.
[摘要] 为了研究软骨形成性肥大的基础机制，我们需要适当的体外间充质干细胞（MSC）的增生模型。该协议描述了我们定义的诱导人脐带血衍生的MSC（hUCB-MSC）的体外软骨形成性肥大的方法。通过将甲状腺激素（T3;三碘甲状腺原氨酸）和最小成骨诱导因子添加到培养基中，我们可以在体外诱导hUCB-MSCs的增生。使用免疫组织化学分析，Western印迹和逆转录酶聚合酶链反应验证肥大性诱导。
关键词：间充质干细胞，体外软骨形成性肥大，成软骨分化，三碘甲腺原氨酸;
[背景] ...
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| Porous Scaffold Seeding and Chondrogenic Differentiation of BMSC-seeded Scaffolds
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Author:
Date:
2015-12-20
[Abstract] Bone marrow-derived mesenchymal stromal stem cells (BMSCs) are a promising cell source for treating articular cartilage defects (Bornes et al., 2014). BMSCs can be seeded within porous biomaterial scaffolds that support three-dimensional cell organization, chondrogenic differentiation and extracellular matrix deposition for the creation of engineered cartilage. This protocol describes our defined methods for isolation and expansion of human and ovine BMSCs, seeding of BMSCs within porous scaffolds and in vitro chondrogenic differentiation (Adesida et al., 2012; Bornes et al., 2015).
[摘要] 骨髓来源的间充质干细胞（BMSCs）是治疗关节软骨缺陷的有希望的细胞来源（Bornes等人，2014）。 BMSCs可以种植在支持三维细胞组织，软骨形成分化和细胞外基质沉积的多孔生物材料支架中，用于创建工程化软骨。 该方案描述了我们定义的用于分离和扩增人和绵羊BMSCs，在多孔支架内接种BMSCs和在体外软骨形成分化的方法（Adesida等人，2012; Bornes et al。，2015）。
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| Mouse ESC Differentiation to Nkx2.1+ Lung and Thyroid Progenitors
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Author:
Date:
2012-11-20
[Abstract] The de novo derivation of lung progenitors from pluripotent stem cells provides the opportunity to model early lung development in vitro and allows easy access to cells for tissue engineering or basic cell biology studies. This detailed protocol allows the generation of lung and thyroid progenitors from mouse embryonic stem cell (ESC) or induced pluripotent stem cell (iPSC) lines. When used together with a published Nkx2.1-GFP knock-in ESC line, the protocol allows tracking and purification of lung and thyroid progenitors by sorting on the GFP reporter based on the induction of the earliest known marker of lung and thyroid cell fate, Nkx2.1. After sorting, a pure population of Nkx2.1+ cells can then be replated for further expansion, differentiation, and maturation in ...
[摘要] 来自多能干细胞的肺祖细胞的新发现提供了在体外模拟早期肺发育的机会，并且容易获得用于组织工程或基础细胞生物学研究的细胞。 该详细方案允许从小鼠胚胎干细胞（ESC）或诱导多能干细胞（iPSC）系生成肺和甲状腺祖细胞。 当与公布的Nkx2.1-GFP敲入ESC系一起使用时，该方案允许通过基于对最早已知的肺和甲状腺细胞命运的标记物的诱导在GFP报告基因上进行分选来跟踪和纯化肺和甲状腺祖细胞， Nkx2.1。 在分选后，然后可以在无血清条件下培养纯化的Nkx2.1 +细胞群，以进一步扩增，分化和成熟。
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